Monoclonal antibody against G3BP: preparation, characterization and its application in analysis of human tumors / 中华病理学杂志
Chinese Journal of Pathology
; (12): 215-219, 2005.
Article
en Zh
| WPRIM
| ID: wpr-265146
Biblioteca responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To better understand the molecular mechanism of tumorigenesis and progression, the monoclonal antibody against G3BP (Ras-GAP SH3 binding protein), which serves as an important downstream effector of Ras signaling, was prepared, characterized and utilized in analysis of various human tumors.</p><p><b>METHODS</b>By using the prokaryotic expression vector pGEX-5X1, GST-G3BP fusion protein was expressed in E. coli BL21 under induction of IPTG. Purified GST-G3BP fusion protein was used to immunize BALB/c mice. The monoclonal antibody against G3BP was produced through conventional hybridoma method and characterized by ELISA, Western blot and immunohistochemical staining.</p><p><b>RESULTS</b>A hybridoma cell line secreting anti-G3BP IgG1 subtype antibody was obtained. Western blot and competitive inhibition assay showed that the antibody was G3BP-specific. Immunohistochemical staining demonstrated that G3BP was over-expressed in formalin-fixed and paraffin-embedded tissues of some human tumors, such as lung cancer, colon cancer, gastric cancer and breast cancer. In breast cancer specimens, the degree of G3BP expression correlated positively with the presence of lymph node metastasis and c-erbB2 expression.</p><p><b>CONCLUSIONS</b>The G3BP-specific monoclonal antibody derived from recombination protein can be used in ELISA, Western blot and immunohistochemical assay. It may provide an important tool in analysis of G3BP in in vitro and in vivo experiments. Besides, G3BP may serve as another prognostic marker for breast cancer.</p>
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Base de datos:
WPRIM
Asunto principal:
Patología
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Proteínas Recombinantes de Fusión
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Neoplasias de la Mama
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Células Tumorales Cultivadas
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Proteínas Portadoras
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Biomarcadores de Tumor
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ADN Helicasas
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Receptor ErbB-2
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Secreciones Corporales
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ARN Helicasas
Límite:
Animals
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Female
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Humans
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Male
Idioma:
Zh
Revista:
Chinese Journal of Pathology
Año:
2005
Tipo del documento:
Article